ES Cell Line Derivations

Requirements from the WUMS investigator are detailed in the "Protocols" section.

1. 10 -/- proven stud males between the ages of 2 - 6 months singly housed for at least 1 week prior to mating
2. 10 -/- designated females between the ages of 3 - 4 weeks
3. Most recent DCM sentinel reports of the rooms housing the above mice

Within the confines of the WUMS investigator's mouse colony, the ES core will:

1. superovulate 10 -/- designated female s and set-up the matings
2. check 0.5 day p.c. plugs

On day 3.5 p.c., investigator will bring his pregnant females to the ES core for blasocyst harvesting.

Culturing the explants for up to 2 weeks in an attempt to isolate ES-like colonies in terms of both morphology and growth pattern.

Culturing may continue for an additional 2 weeks until a preliminary freeze is made possible.

The ES core can make arrangements for mycoplasma, MAP, and karyotyping if requested.

Primary expansions for up to 30 vials for each early passage ES cell line. This can take anywhere from 10 - 14 days.